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Stan and Kyle are obsessed with the video game Guitar Hero, and attempt to break the game's scoring record in front of their friends. As soon as they break the 100,000 points record on "Carry on Wayward Son", they are contacted by Charles Kincaid, a talent agent, who wants to offer them recording contracts. They become Guitar Hero rock star celebrities overnight, enjoying the hedonistic "sex and drugs" lifestyle. The record company organizes a future event where Stan and Kyle will attempt to break a 1 million point record in front of a live audience. Stan is taken aside by his manager and told that Kyle is holding him back, and encourages Stan to dump Kyle in favor of another kid, Thad Jarvis, who can play many Guitar Hero songs "acoustically", having memorized what buttons to press instead of relying on the on-screen cues.

Kyle discovers Stan practicing with Thad and angrily breaks up their "band". Kyle goes on to perform at a local bowling alley by its owner for free food and drink, improving as he plays. Stan becomes stressed out over the upcoming event, and tries to seek refuge using a different video game Heroin Hero where the only objective is literally chasing a dragon. The game somehow shortens his temper and concentration into getting into a fight with Thad, who subsequently leaves Stan. Stan, still intoxicated with Heroin Hero, is forced to play the event solo, ending up getting a miserable score and throwing up on stage.

Stan comes back to the video game store and decides to pick another game to play, but when offered by the game store owner Rehab Hero (which remedies the heroin-like addiction to a former player of "Heroin Hero"), Stan instead selects a simple driving game. While driving around in-game, Stan hears "Carry on Wayward Son" from the game's soundtrack and remembers the good time he had with Kyle. Stan goes to apologize to Kyle and after a brief spat, the two become friends again, dumping the recording contract. They go back home and go on to break the 1 million point record in front of their friends. To their shock, the game rewards them by telling them, "Congratulations You Are Fags!". The two become upset that they have worked so hard only to be insulted, and angrily walk out; Cartman and Butters start to play the game, with Butters eagerly requesting that he gets to be the one to betray Cartman "after the sex and drugs party".

Jay Cutler, the then-quarterback for the Denver Broncos, was insulted in the episode ("You kind of suck, but my dad says you might be good some day"), and responded lightheartedly, saying "I thought it was funny. They can make fun of me if they want to."[5]

When Stan's new Guitar Hero partner says "I quit! I quit, I quit, I quit!" is a reference to the film That Thing You Do!.[6]

The manager Charles Kincaid is a reference to Reuben Kincaid of The Partridge Family and even looks like him.

The episode drew in 4 million total viewers (P2+) and was the most watched telecast of the year on Comedy Central, as well as the most watched program in all of television for Wednesday night among men 18–34 and the most watched program on cable among persons 18–49.[7] The episode was the highest rating since "Cartman's Mom Is Still a Dirty Slut",[8] beating the previous record-holder "Imaginationland Episode III", the episode that aired one week before "Guitar Queer-o".[9]

Despite good ratings, the episode received some mediocre reviews. IGN called it a "lackluster episode" saying that it was "based on a concept that could have proved fun – but ultimately doesn't seem to be enough for an entire episode."[10] 411Mania also criticized the episode, saying that "the first twenty-one and a half minutes weren't funny at all" claiming that the last line of the episode was the best part.[11]TV Squad, on the other hand, gave the episode a somewhat positive review.[12]

A portion of the episode was used on NPR's Day to Day to discuss the idea that the popularity and interest in video games like Guitar Hero III or Rock Band could supplant traditional instruments for kids.[13]

The New York Times identifed Guitar Hero's reference in the episode as the most effective TV product placement for that week.[14][15]

"Guitar Queer-O", along with the thirteen other episodes from South Park's eleventh season, were released on a three-disc DVD set in the United States on August 12, 2008. The set includes brief audio commentaries by Parker and Stone for each episode.[16]